Sulfate Molecules Research Articles

Neuroectodermal Stem Cells Grafted into the Injured Spinal Cord Induce Both Axonal Regeneration and Morphological Restoration via Multiple Mechanisms.

Spinal cord contusion injury leads to severe loss of gray and white matter and subsequent deficit of motor and sensory functions below the lesion. In this study, we investigated whether application of murine clonal embryonic neuroectodermal stem cells can prevent the spinal cord secondary damage and induce functional recovery. Stem cells (NE-GFP-4C cell line) were grafted intraspinally or intravenously immediately or one week after thoracic spinal cord contusion injury. Control animals received cell culture medium or fibrin intraspinally one week after injury. Functional tests (Basso, Beattie, Bresnahan, CatWalk®) and detailed morphological analysis were performed to evaluate the effects of grafted cells. Stem cells applied either locally or intravenously induced significantly improved functional recovery compared with their controls. Morphologically, stem cell grafting prevented the formation of secondary injury and promoted sparing of the gray and white matters. The transplanted cells integrated into the host tissue and differentiated into neurons, astrocytes, and oligodendrocytes. In intraspinally grafted animals, the corticospinal tract axons regenerated along the ventral border of the cavity and have grown several millimeters, even beyond the caudal end of the lesion. The extent of regeneration and functional improvement was inversely related to the amounts of chondroitin sulphate and ephrin-B2 molecules around the cavity and to the microglial and astrocytic reactions in the injured segment early after injury. The grafts produced glial cell derived neurotrophic factor, macrophage inflammatory protein-1a, interleukin (IL)-6 and IL-10 in a paracrine fashion for at least one week. Treating the grafted cords with neutralizing antibodies against these four factors through the use of osmotic pumps nearly completely abolished the effect of the graft. The non-significant functional improvement after function blocking is likely because the stem cell derivatives settled in the injured cord. These data suggest that grafted neuroectodermal stem cells are able to prevent the secondary spinal cord damage and induce significant regeneration via multiple mechanisms.

Mapping of a Regulatory Site of the Escherichia coli ADP-Glucose Pyrophosphorylase.

The enzyme ADP-glucose pyrophosphorylase (ADP-Glc PPase) controls the biosynthesis of glycogen in bacteria and starch in plants. It is regulated by various activators in different organisms according to their metabolic characteristics. In Escherichia coli, the major allosteric activator is fructose 1,6-bisphosphate (FBP). Other potent activator analogs include 1,6-hexanediol bisphosphate (HBP) and pyridoxal 5′-phosphate (PLP). Recently, a crystal structure with FBP bound was reported (PDB ID: 5L6S). However, it is possible that the FBP site found is not directly responsible for the activation of the enzyme. We hypothesized FBP activates by binding one of its phosphate groups to another site (“P1”) in which a sulfate molecule was observed. In the E. coli enzyme, Arg40, Arg52, and Arg386 are part of this “P1” pocket and tightly complex this sulfate, which is also present in the crystal structures of ADP-Glc PPases from Agrobacterium tumefaciens and Solanum tuberosum. To test this hypothesis, we modeled alternative binding conformations of FBP, HBP, and PLP into “P1.” In addition, we performed a scanning mutagenesis of Arg residues near potential phosphate binding sites (“P1,” “P2,” “P3”). We found that Arg40 and Arg52 are essential for FBP and PLP binding and activation. In addition, mutation of Arg386 to Ala decreased the apparent affinity for the activators more than 35-fold. We propose that the activator binds at this “P1” pocket, as well as “P2.” Arg40 and Arg52 are highly conserved residues and they may be a common feature to complex the phosphate moiety of different sugar phosphate activators in the ADP-Glc PPase family.

Sodium dodecyl sulfate decorated Legionella pneumophila for enhanced detection with a GaAs/AlGaAs nanoheterostructure biosensor

The net electric charge associated with a bacterial strain is primarily defined by the number of available functional groups at its surface and we observed that it can determine the limit of detection of a charge-sensing biosensor. We have investigated the dynamic range of bacterial electric charge variations through binding negatively charged sodium dodecyl sulphate (SDS) molecules, with the objective of improving the detection limit of a charge-sensing GaAs/AlGaAs nanoheterostructure biosensor designed for detection of Legionella pneumophila. A two-fold increased zeta potential of L. pneumophila was measured at pH 7.4 following the exposure of these bacteria to an SDS solution at 0.02 mg/mL. Subsequently, it was possible to detect SDS decorated and heat-inactivated L. pneumophila at 103 CFU/mL. This illustrates the fundamental role of the bacterial electric charge in the operation of photocorrosion-based III-V semiconductor biochips. We discuss the mechanisms of bacterial interaction with SDS, critical aspects of decorating bacteria with this anionic surfactant and the channels responsible for charge transfer.

Sulfated glycosaminoglycans and low-density lipoprotein receptor contribute to Clostridium difficile toxin A entry into cells.

Clostridium difficile toxin A (TcdA) is a major exotoxin contributing to disruption of the colonic epithelium during C. difficile infection. TcdA contains a carbohydrate-binding CROPs (combined repetitive oligopeptides) domain that mediates its attachment to cell surfaces, but recent data suggest the existence of CROPs-independent receptors. Here we carried out genome-wide CRISPR-Cas9-mediated screens using a truncated TcdA lacking the CROPs and identified sulfated glycosaminoglycans (sGAGs) and low-density lipoprotein receptor (LDLR) as host factors contributing to binding and entry of TcdA. TcdA recognizes the sulfation group in sGAGs. Blocking sulfation and GAG synthesis reduces TcdA binding and entry into cells. Binding of TcdA to the colonic epithelium can be reduced by surfen, a small molecule that masks sGAGs, by GM-1111, a sulfated heparan sulfate analog, and by sulfated cyclodextrin, a sulfated small molecule. Cells lacking LDLR also show reduced sensitivity to TcdA, although binding between LDLR and TcdA are not detected, suggesting that LDLR may facilitate endocytosis of TcdA. Finally, GM-1111 reduces TcdA-induced fluid accumulation and tissue damage in the colon in a mouse model of injecting TcdA into the cecum. These data demonstrate in vivo and pathological relevance of TcdA-sGAGs interactions, and reveal a potential therapeutic approach of protecting colonic tissues by blocking TcdA-sGAGs interactions.

Polydispersity and negative charge are key modulators of extracellular matrix deposition under macromolecular crowding conditions

Macromolecular crowding is a biophysical phenomenon that stems from the volume excluded by macromolecules, as they undergo steric repulsion and electrostatic interactions. The excluded volume depends on the shape, size, charge and polydispersity of the molecules. Although theoretical/computational models have been used to assess the influence of macromolecular crowding in biological media, real-time experiments are scarce. Herein, we evaluated the influence of hydrodynamic radius, charge and polydispersity of (a) various concentrations of different crowders (carrageenan, Ficoll™ and dextran sulphate); (b) various molecular weights of different crowders (70, 400 and 100 kDa of Ficoll™ and 10, 100 and 500 kDa of dextran sulphate) and (c) various cocktails of the same crowders (cocktails of various concentrations of different molecular weights Ficoll™ and dextran sulphate) on extracellular matrix deposition in human dermal fibroblast culture. The use of crowding cocktails with different molecular weight/concentrations of Ficoll™ or dextran sulphate molecules led to increased polydispersity and enhanced collagen type I deposition in comparison to their mono-domain counterparts. Carrageenan, however, induced the highest deposition of collagen type I due to its negative charge and inherent polydispersity. Our data contribute to a better understanding of the influence of the biophysical properties of the crowders on extracellular matrix deposition in vitro. Statement of SignificanceMacromolecular crowding is a biophysical phenomenon that accelerates and enhances extracellular matrix deposition in cell culture systems. Herein, we demonstrate that negatively charged and polydispersed macromolecules or cocktails of macromolecules, as opposed to neutral and monodomain macromolecules, induce highest extracellular matrix deposition in human dermal fibroblast cultures.

Sulfation made simple: a strategy for synthesising sulfated molecules.

The study of organosulfates is a burgeoning area in biology, yet there are significant challenges with their synthesis. We report the development of a tributylsulfoammonium betaine as a high yielding route to organosulfates. The optimised reaction conditions were interrogated with a diverse range of alcohols, including natural products and amino acids.

The results of postgenomic analysis of a glucosamine sulfate molecule indicate the prospects of treatment for comorbidities

Postgenomic analysis of the effects of active ingredients of drugs involves the evaluation of the effects of relevant molecules on the transcription of genes (transcriptomes), on the changes in the activity of proteins (proteomes), and on the activity of molecular cascades (reactomes). The paper gives the results of applying the current chemoinformational approaches to the postgenomic analysis of the effects of glucosamine sulfate (GS). The main result of the investigation is to simultaneously establish the synergistic effect of GS on transcriptomes, proteomes, and reactomes. In particular, GS assists in reducing not only the transcription of genes involved in the NF-κB proinflammatory signaling cascade (NFKB2, TNFRSF1B, PYCARD, TRAF2, TNFSF12, etc.), but also the activity of proteomic proteins that transmit a signal at different levels of the NF-κB cascade (CD44, TLR4, ICAM1, NF-κB, JAK/STAT, etc.). The complex anti-inflammatory effect of GS in reducing the synthesis of proinflammatory cytokines and weakening their effects on the cells is pathogenetic in the treatment of not only osteoarthritis, but also comorbidities accompanied by chronic inflammation.

Palladium nanoparticles from surfactant/fast-reduction combination one-pot synthesis for the liquid fuel cell applications

We report a facile one-pot synthesis of well-dispersed Pd nanoparticles on a carbon nanofiber (CNF) support, achieved via the unique sequential processes of supramolecular self-assembly and fast reduction of the Pd source. Pd ions attached to the sulfate ends of sodium dodecyl sulfate (SDS) molecules self-assembled on a CNF, and subsequent fast reduction suppressed particle coarsening, in contrast to the slow reduction processes in previous works. The particle size and dispersion uniformity were comparable to those of particles synthesized by combined SDS/high-energy irradiation. The Pd nanoparticles prepared using the present one-pot approach were superior or comparable to those obtained using non-one-pot approaches in previous works in terms of electrocatalytic activity and long-term stability in the electrooxidation of liquid fuels at the anodes of direct formic acid, direct methanol, and direct ethanol fuel cells.

Heparan sulfate chain length modulates the cellular targeting of prions in vivo

Prion diseases are fatal neurodegenerative disorders caused by the misfolding of the physiologically expressed prion protein into an insoluble form that accumulates as amorphous aggregates (subfibrillar prions) or amyloid fibers (fibrillar prions). Identical prion protein sequences can misfold into different conformational variants or strains that affect specific neural cell populations, leading to a wide variety of clinical signs and brain regions targeted. The mechanisms underlying the cell targeting of prion strains are unknown. Heparan sulfate (HS) molecules are glycosaminoglycans abundantly expressed in the brain and that profoundly vary in length and sulfation depending on the cell type. HS co‐localize with prion plaques of Creutzfeldt–Jakob disease (CJD), Gerstmann‐Sträussler‐Scheinker (GSS) syndrome, and sheep scrapie, and promote prion replication and fibril formation in vitro. Paradoxically, HS have been used to slow or even prevent prion formation in CJD patients and scrapie‐infected rodents. We hypothesize that the interaction of HS with prions is a major determinant underlying the selective cell vulnerability in prion diseases. Here we characterized the HS length that most effectively modulates the in vitro prion conversion into the aggregated state. We also determined how manipulating the HS chain length shifts the cell tropism of prion strains in vivo. While longer HS chain lengths led to more efficient prion conversion for subfibrillar strains (RML and ME7), intermediate lengths were more effective for the fibrillar strain mCWD. Ext1+/‐ mice expressing shorter HS molecules only altered prion disease phenotype when inoculated with the fibrillar prion mCWD, affecting the disease progression and neuropathology. Together these results suggest that the specific length of the HS together with the prion aggregate conformation determine the binding affinity of prions to HS, and underlie the specific cellular targeting in the CNS. These results have implications for other misfolding proteins such as amyloid‐β and α‐synuclein, which also interact with HS.Support or Funding InformationNIH, NINDS R01 grant 069566 Creutzfeldt–Jakob disease Foundation Fundación Ramón ArecesThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Ultrasound Effect on Molecules of Sodium Dodecyl Sulphate as Systems of Nanoparticles

Ultrasound Effect on Molecules of Sodium Dodecyl Sulphate as Systems of Nanoparticles

Electrochemical oxidation of As(iii) on Pd immobilized Pt surface: kinetics and sensing performance.

Pd nanoparticles were electrochemically immobilized on a Pt surface in the presence of sodium dodecyl sulfate (SDS) molecules to study the electrokinetics of arsenite oxidation reactions and the corresponding sensing activities. The X-ray photoelectron spectroscopy (XPS) analysis showed that on the Pt surface, Pd atoms exist as adatoms and the contents of Pd(0) and Pd(ii) were 75.72 and 24.28 at%, respectively, and the particle sizes were in the range of 61–145 nm. The experimental results revealed that the catalytic efficiency as well as the charge transfer resistance (at the redox potential of the Fe(ii)/Fe(iii) couple) increased in the order of Pt < Pt–Pd < Pt–Pdsds. A Pt–Pdsds electrode exhibited an open circuit potential (OCP) of 0.65 V in acidic conditions; however, when 50.0 mM NaAsO2 was present, the OCP value shifted to 0.42 V. It has been projected that the As(iii) oxidation proceeds using a sequential pathway: As(iii) → As(iv) → As(v). After optimization of the square wave voltammetric data, the limits of detection of As(iii) were obtained as 1.3 μg L−1 and 0.2 μg L−1 when the surface modification of the Pt surface was executed with Pd particles in the absence and presence of the SDS surfactant, respectively. Finally, real samples were analyzed with excellent recovery performance.

Surfactant and heavy metal interaction in poplar: a focus on SDS and Zn uptake.

Surfactants are widely used detergent ingredients and, thanks to their chemical properties, they are applied for remediation of sites polluted by heavy metals and organic contaminants, both in soil flushing and in phytoremediation. However, their direct effects on tree physiology especially in consociation with heavy metal pollution, as well as their possible absorption by plants, have not been appropriately investigated. In order to evaluate plant uptake/translocation of the surfactant sodium dodecyl sulfate (SDS) and the heavy metal zinc (Zn) in Populus alba L. Villafranca clone, SDS was applied alone (0.5 mM) or in combination with Zn (1 mM). Physiological effects on plant growth and photosynthetic performance were investigated. An increasing trend of Zn translocation towards basal leaves as a consequence of SDS co-treatment (1 mM Zn + 0.5 mM SDS; P = 0.03) was observed, proving the ability of SDS to improve heavy metals translocation. However, SDS exposure (both in 0.5 mM SDS and 1 mM Zn + 0.5 mM SDS treated plants) resulted in the appearance of foliar necrosis that expanded with an acropetal trend and finally led to leaf abscission. This phenotype may be caused by the emergence of an additional stress during the experimental trial, which could be related to the dissociation of sodium (Na) ions from the dodecyl sulfate molecules in the hydroponic system. In fact, while liquid chromatography-tandem mass spectrometry measurements revealed that dodecyl sulfate is mainly retained at root levels, Na is translocated to the aerial parts of the plant.

Adsorption kinetics of retarding admixtures on cement with time controlled addition

Retarders are widely used in construction materials to control the setting and hardening. The present study focuses on changes in the rheological behavior, the admixture adsorption and the pore solution composition with a controlled addition of sodium gluconate and ethylenediamine tetra(methylene phosphonic acid) (EDTMP) in cement.The adsorption of the sodium gluconate was influenced by time of addition of dosage. Delayed addition induced a depot effect which was reduced by the ongoing hydration whereas the adsorption of EDTMP was not affected by time of dosage. Nearly the complete admixture concentration was directly adsorbed on clinker phases or first hydration products.For sodium gluconate, an interaction with ferrite containing clinker phases and a calcium complexing mechanism were suggested. In contrast, EDTMP seemed to have no significant influence on the calcium concentration. With regard to the sodium concentration a fast adsorption of sodium gluconate was determined. Furthermore, a competitive adsorption between dissolved sulfate and retarder molecules or a complex formation mechanism is supposed.

The Sea as a Rich Source of Structurally Unique Glycosaminoglycans and Mimetics.

Glycosaminoglycans (GAGs) are sulfated glycans capable of regulating various biological and medical functions. Heparin, heparan sulfate, chondroitin sulfate, dermatan sulfate, keratan sulfate and hyaluronan are the principal classes of GAGs found in animals. Although GAGs are all composed of disaccharide repeating building blocks, the sulfation patterns and the composing alternating monosaccharides vary among classes. Interestingly, GAGs from marine organisms can present structures clearly distinct from terrestrial animals even considering the same class of GAG. The holothurian fucosylated chondroitin sulfate, the dermatan sulfates with distinct sulfation patterns extracted from ascidian species, the sulfated glucuronic acid-containing heparan sulfate isolated from the gastropode Nodipecten nodosum, and the hybrid heparin/heparan sulfate molecule obtained from the shrimp Litopenaeus vannamei are some typical examples. Besides being a rich source of structurally unique GAGs, the sea is also a wealthy environment of GAG-resembling sulfated glycans. Examples of these mimetics are the sulfated fucans and sulfated galactans found in brown, red and green algae, sea urchins and sea cucumbers. For adequate visualization, representations of all discussed molecules are given in both Haworth projections and 3D models.

Chondroitin Sulfate–Based Biocompatible Crosslinker Restores Corneal Mechanics and Collagen Alignment

To evaluate the crosslinking effect of functionalized chondroitin sulfate (CS) in an ex vivo rabbit cornea model. Chondroitin sulfate molecules were chemically modified with the N-hydroxysuccinimide (NHS) group. Enucleated rabbit eyes were crosslinked with 2, 5, or 10 mg/mL CS-NHS solution for 30 or 60 minutes. The CS-NHS penetration, corneal swelling ratio, Young's modulus, and ultrastructure of the crosslinked corneas were characterized. In addition, rabbit corneas were further treated with a collagenase-chondroitinase solution to create an ex vivo keratoconus (KC)-like model. The KC model corneas were crosslinked with a standard riboflavin-ultraviolet (UV) method or alternatively with CS-NHS. Corneal mechanics, ultrastructure, and keratocyte gene expression were evaluated after UV and CS-NHS crosslinking. CS-NHS effectively penetrated into the corneal stroma within 60 minutes of treatment initiation. CS-NHS crosslinking reduced the swelling ratio by 35%, increased Young's modulus by 20%, and increased collagen fibril diameter and density. CS-NHS crosslinking improved corneal mechanics of KC model corneas to levels comparable to those with UV crosslinking. Moreover, CS-NHS crosslinking demonstrated significant downregulation of proinflammatory gene expression of keratocytes, indicating a potential protective effect imparted by CS-NHS during crosslinking. Our results demonstrated that CS-NHS can reinforce normal and KC model corneal mechanics, and restore collagen density and alignment in KC model corneas without causing extensive keratocyte apoptosis and proinflammatory gene upregulation. Therefore, CS-NHS crosslinking can potentially provide an effective, safe, and biocompatible means of corneal reinforcement.

Heterologous expression in Pichia pastoris and biochemical characterization of the unmodified sulfatase from Fusarium proliferatum LE1.

Sulfatases are a family of enzymes (sulfuric ester hydrolases, EC 3.1.6.-) that catalyze the hydrolysis of a wide array of sulfate esters. To date, despite the discovery of many sulfatase genes and the accumulation of data on numerous sulfated molecules, the number of characterized enzymes that are key players in sulfur metabolism remains extremely limited. While mammalian sulfatases are well studied due to their involvement in a wide range of normal and pathological biological processes, lower eukaryotic sulfatases, especially fungal sulfatases, have not been thoroughly investigated at the biochemical and structural level. In this paper, we describe the molecular cloning of Fusarium proliferatum sulfatase (F.p.Sulf-6His), its recombinant expression in Pichia pastoris as a soluble and active cytosolic enzyme and its detailed characterization. Gel filtration and native electrophoretic experiments showed that this recombinant enzyme exists as a tetramer in solution. The enzyme is thermo-sensitive, with an optimal temperature of 25°C. The optimal pH value for the hydrolysis of sulfate esters and stability of the enzyme was 6.0. Despite the absence of the post-translational modification of cysteine into Cα-formylglycine, the recombinant F.p.Sulf-6His has remarkably stable catalytic activity against p-nitrophenol sulfate, with kcat = 0.28 s-1 and Km = 2.45 mM, which indicates potential use in the desulfating processes. The currently proposed enzymatic mechanisms of sulfate ester hydrolysis do not explain the appearance of catalytic activity for the unmodified enzyme. According to the available models, the unmodified enzyme is not able to perform multiple catalytic acts; therefore, the enzymatic mechanism of sulfate esters hydrolysis remains to be fully elucidated.

Abstract 946: Characterize and identify secreted high molecular weight heparanase from PC3M conditioned medium

Abstract Introduction: Neutrophils, platelets, lymphocytes are typical sources of NAP-2 which is associated with heparanase activity. Heparanase degrades polymeric heparan sulfate molecules into shorter oligosaccharides, participating in extracellular matrix degradation. It is active under acidic conditions during tumor invasion and inflammatory processes. Our lab previously found that some cell lines like androgen independent prostate cancer cells (PC3M) can secrete a protein that reacts with anti-NAP-2 antibody, but has a much higher molecular weight(&amp;gt;50KD) than NAP-2 (MW~6-14 KD), and has heparanase activity. The purpose of this study is to characterize and identify this protein. Procedures: Immunoprecipitation was performed using Pierce DirectIP kit according the manufacturer’s instructions. Heparin binding affinity column used HiTrap™ Heparin HP column from GE Healthcare Bio-Sciences. The binding buffer was 10mM NaAcetate pH 5.0, the eluting buffers were 0.5M-2M NaCl in 10mM NaAcetate pH 5.0. The mass spectrometry was done by KCI Proteomics Core facility. Data: The immunoprecipitation method confirmed that this higher molecular weight protein indeed can be pulled down by the NAP-2 antibody, and after using heparin binding affinity column to purify, we can still get the &amp;gt; 50KD band on the western blot for NAP-2, meaning it is a heparin binding protein. Conclusions: A protein identified with NAP-2 antibody, but with higher molecular weight than native NAP-2 is found in the conditioned medium of PC3M cells that binds heparin, further confirming a potential role in tumor progression for prostate cancer. Citation Format: Donghong Ju, Mary A. Kosir. Characterize and identify secreted high molecular weight heparanase from PC3M conditioned medium [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 946. doi:10.1158/1538-7445.AM2017-946

Heterologous expression in Pichia pastoris and biochemical characterization of the unmodified sulfatase from Fusarium proliferatum LE1.

Sulfatases are a family of enzymes (sulfuric ester hydrolases, EC 3.1.6.-) that catalyze the hydrolysis of a wide array of sulfate esters. To date, despite the discovery of many sulfatase genes and the accumulation of data on numerous sulfated molecules, the number of characterized enzymes that are key players in sulfur metabolism remains extremely limited. While mammalian sulfatases are well studied due to their involvement in a wide range of normal and pathological biological processes, lower eukaryotic sulfatases, especially fungal sulfatases, have not been thoroughly investigated at the biochemical and structural level. In this paper, we describe the molecular cloning of Fusarium proliferatum sulfatase (F.p.Sulf-6His), its recombinant expression in Pichia pastoris as a soluble and active cytosolic enzyme and its detailed characterization. Gel filtration and native electrophoretic experiments showed that this recombinant enzyme exists as a tetramer in solution. The enzyme is thermo-sensitive, with an optimal temperature of 25°C. The optimal pH value for the hydrolysis of sulfate esters and stability of the enzyme was 6.0. Despite the absence of the post-translational modification of cysteine into Cα-formylglycine, the recombinant F.p.Sulf-6His has remarkably stable catalytic activity against p-nitrophenol sulfate, with kcat = 0.28 s-1 and Km = 2.45 mM, which indicates potential use in the desulfating processes. The currently proposed enzymatic mechanisms of sulfate ester hydrolysis do not explain the appearance of catalytic activity for the unmodified enzyme. According to the available models, the unmodified enzyme is not able to perform multiple catalytic acts; therefore, the enzymatic mechanism of sulfate esters hydrolysis remains to be fully elucidated.

Magnetization transfer in a partly deuterated lyotropic liquid crystal by single- and dual-frequency RF irradiations

The mechanism of magnetization transfer (MT) in a lyotropic liquid crystal made of sodium dodecyl sulfate, decanol, and water molecules is investigated by using deuterated molecules and single- and dual-frequency RF irradiations. The resulting Z-spectra suggest that the decanol molecules are mainly responsible for the MT effects in this system, through proton exchange to water. This is further confirmed by monitoring the relaxation of dipolar order, which allows one to estimate the transfer rate of magnetization from decanol to water. The potential benefits of using dual-frequency RF irradiation for inducing MT effects are explored through numerical solutions to a MT model based on Provotorov’s partial saturation theory.

A molecular dynamics-based algorithm for evaluating the glycosaminoglycan mimicking potential of synthetic, homogenous, sulfated small molecules.

Glycosaminoglycans (GAGs) are key natural biopolymers that exhibit a range of biological functions including growth and differentiation. Despite this multiplicity of function, natural GAG sequences have not yielded drugs because of problems of heterogeneity and synthesis. Recently, several homogenous non-saccharide glycosaminoglycan mimetics (NSGMs) have been reported as agents displaying major therapeutic promise. Yet, it remains unclear whether sulfated NSGMs structurally mimic sulfated GAGs. To address this, we developed a three-step molecular dynamics (MD)-based algorithm to compare sulfated NSGMs with GAGs. In the first step of this algorithm, parameters related to the range of conformations sampled by the two highly sulfated molecules as free entities in water were compared. The second step compared identity of binding site geometries and the final step evaluated comparable dynamics and interactions in the protein-bound state. Using a test case of interactions with fibroblast growth factor-related proteins, we show that this three-step algorithm effectively predicts the GAG structure mimicking property of NSGMs. Specifically, we show that two unique dimeric NSGMs mimic hexameric GAG sequences in the protein-bound state. In contrast, closely related monomeric and trimeric NSGMs do not mimic GAG in either the free or bound states. These results correspond well with the functional properties of NSGMs. The results show for the first time that appropriately designed sulfated NSGMs can be good structural mimetics of GAGs and the incorporation of a MD-based strategy at the NSGM library screening stage can identify promising mimetics of targeted GAG sequences.